Use of reference gene expression in rat distal colon after radiation exposure: A caveat
| Autor: | Anna Ropenga, N. M. Griffiths, Marie Vandamme, Alain Chapel |
|---|---|
| Přispěvatelé: | Radiobiologie et épidémiologie (DRPH/SRBE), Institut de Radioprotection et de Sûreté Nucléaire (IRSN) |
| Jazyk: | angličtina |
| Rok vydání: | 2004 |
| Předmět: |
serotonin 1F receptor
Male [SDV]Life Sciences [q-bio] tissue distribution Messenger Wistar complementary DNA radiation exposure animal cell Wistar rat Transcriptome 0302 clinical medicine Transcription (biology) Reference Values rat animal genetics acidic ribosomal protein P0 comparative study housekeeping gene 0303 health sciences Radiation messenger RNA Reverse Transcriptase Polymerase Chain Reaction article artifact standard methodology gene expression regulation 3. Good health Housekeeping gene unclassified drug Reverse transcription polymerase chain reaction priority journal Hypoxanthine-guanine phosphoribosyltransferase 030220 oncology & carcinogenesis ionizing radiation radiation dose Artifacts cyclophilin A radiation response Colon Metabolic Clearance Rate animal experiment DNA transcription Biophysics Biology Radiation Dosage animal tissue glyceraldehyde 3 phosphate dehydrogenase reverse transcription polymerase chain reaction Dose-Response Relationship 03 medical and health sciences Ribosomal protein Complementary DNA gene expression profiling Animals Radiology Nuclear Medicine and imaging controlled study RNA Messenger Rats Wistar Gene 030304 developmental biology hypoxanthine phosphoribosyltransferase nonhuman animal model reference value Dose-Response Relationship Radiation Molecular biology Rats descending colon physiology gene expression RNA transcriptome metabolism |
| Zdroj: | Radiation Research Radiation Research, 2004, 161 (5), pp.597-602. ⟨10.1667/RR3173⟩ |
| DOI: | 10.1667/RR3173⟩ |
| Popis: | Research on the effects of ionizing radiation exposure includes transcriptome studies using real-time reverse transcription polymerase chain reaction (RT-PCR). These studies require the use of a reference gene that normalizes for cDNA quantity and corrects for transcription between different samples. In this study, several criteria are reviewed that allow the choice of a reference gene. With the example of five genes selected from the widely used standard housekeeping genes, Gapd (glyceraldehyde-3-phosphate dehydrogenase), Hprt (hypoxanthine-guanine phosphoribosyl transferase), cyclophilin A, AcRP0 (acidic ribosomal protein P0) and 18S, we show that the use of a reference gene without a preliminary study is hazardous. We have shown in rat colon after a hemi-body irradiation that expression of a gene of interest, the serotonin receptor type IF (5-HT1F), was either increased or unchanged, with the result depending on the reference gene used. This work has led us to propose the use of two reference genes, a ribosomal gene, 18S, and another gene with a level of expression closer to that of the gene of interest. The methodology reported here may be applied to other studies of gene expression levels to evaluate the effects of experimental treatment on the expression of potential reference genes. © 2004 by Radiation Research Society. |
| Databáze: | OpenAIRE |
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